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ObjectiveTo investigate the effect of hypoxia-inducible factor-1α (HIF-1α) on the stemness and epirubicin sensitivity of hepatoma cells. MethodsHepatoma cells were selected for experiment. HepG2 hepatoma cells transfected with HIF-1α overexpression plasmid were selected as experimental group, and those transfected with pcDNA3.1 empty plasmid were selected as control group; HepG2 cells alone were selected as HepG2 group. Quantitative real-time PCR was used to measure the mRNA expression of HIF-1α; Western blot was used to measure the protein expression of HIF-1α; flow cytometry was used to measure the expression of CD133 on the surface of hepatoma cells. The three groups of cells were treated with epirubicin at different concentrations (0, 6.25, 12.5, 25, and 50 μmol/L) for 24 hours; MTT assay was used to measure cell viability, and flow cytometry was used to measure apoptosis after treatment with epirubicin (50 μmol/L). A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the t-test was used for further comparison between two groups. ResultsCompared with the HepG2 group and the control group, the experimental group had a significant increase in the mRNA expression of HIF-1α (both P＜0.001), and Western blot showed high expression of HIF-1α in the experimental group. The percentage of CD133 cells was 0.040%±0.003% in the HepG2 group, 0.030%±0.010% in the control group, and 20.110%±0.600% in the experimental group, and the experimental group had a significantly higher positive rate of CD133+ than the HepG2 group and the control group (both P＜0.001). At an epirubicin concentration of 25 and 50 μmol/L, the HepG2 group and the control group had significantly inhibited cell viability and a significantly lower cell viability than the experimental group (both P＜005). After the treatment with 50 μmol/L epirubicin for 48 hours, the experimental group had a significantly lower cell apoptosis rate than the HepG2 group (67.9%±2.5% vs 93.6%±1.5%, P＜0.001) and the control group (67.9%±2.5% vs 93.0%±1.2%, P＜0001). ConclusionHepG2 cells are successfully transfected with HIF-1α overexpression plasmid, and HIF-1α can increase the percentage of liver cancer stem cells and improve their resistance to epirubicin.
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OBJECTIVES@#Since the outbreak of coronavirus disease 2019 (COVID-19), it has spread rapidly in China and many other countries. The rapid increase in the number of cases has caused widespread panic among people and has become the main public health problem in the world. Severe patients often have difficult breathing and/or hypoxemia after 1 week of onset. A few critically ill patients may not only rapidly develop into acute respiratory distress syndrome, but also may cause coagulopathy, as well as multiple organs failure (such as heart, liver and kidney) or even death. This article is to analyze the predictive role of clinical features in patients with COVID-19 for severe disease, so as to help doctor monitor the severity-related features, restrain the disease progress, and provide a reference for improvement of medical treatment.@*METHODS@#The clinical data of 208 patients with COVID-19 who were isolated and treated in Changsha Public Health Treatment Center from January 17, 2020 to March 14, 2020 were collected. All patients were the mild and ordinary adult patients on admission, including 105 males and 103 females from 19 to 84 (median age 44) years old. According to the "Program for the diagnosis and treatment of novel coronavirus (COVID-19) infected pneumonia (Trial version 7)" issued by the General Office of National Health Committee and Office of State Administration of Traditional Chinese Medicine as the diagnostic and typing criteria. According to progression from mild to severe disease during hospitalization, the patients were divided into a mild group (=183) and a severe transformation group (=25). The clinical features such as age, underlying disease, blood routine, coagulation function, blood biochemistry, oxygenation index, and so on were analyzed. Among them, laboratory tests included white blood cell (WBC), lymphocytes (LYM), neutrophil (NEU), hemoglobin (Hb), platelet (PLT), prothrombin time (PT), plasma fibrinogen (Fib), activated partial prothrombin time (APTT), thrombin time (TT), -dimer, total bilirubin (TBIL), albumin (ALB), alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), serum creatinine (Cr), creatine kinase (CK), creatine kinase isoenzyme-MB (CK-MB), lactate dehydrogenase (LDH), C-reactive protein (CRP), and oxygen partial pressure in arterial blood. Partial pressure of oxygen in arterial blood/fractional concentration of inspiratory oxygen (PaO/FiO) was calculated. The variables with statistical significance were analyzed by logistic regression analysis.@*RESULTS@#Patients in the severe transformation group had more combined underlying diseases than those in the mild group (<0.05). From the perspective of disease distribution, patients in the severe transformation group had more combined hypertension (<0.05). In the severe transformation group, PT was significantly longer, the levels of Fib, ALT, AST, CK, LDH, and CRP were significantly higher than those in the mild group (<0.05 or <0.001), while LYM, ALB, and PaO/FiO were significantly lower than those in the mild group (<0.05 or <0.001). Logistic regression analysis was performed on clinical features with statistically significant differences. Combined with hypertension, LYM, PT, Fib, ALB, ALT, AST, CK, LDH, and CRP as independent variables, and having severe disease or not was the dependent variable. The results show that combined hypertension, decreased LYM, longer PT, and increased CK level were independent risk factors that affected the severity of COVID-19 (<0.05).@*CONCLUSIONS@#The patients with mild COVID-19 who are apt to develop severe diseases may be related to combined hypertension, decreased LYM, and longer PT, and increased CK level. For the mild patients with these clinical features, early intervention may effectively prevent the progression to severe diseases.
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Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Young Adult , Betacoronavirus , China , Coronavirus Infections , Diagnosis , Disease Progression , Hospitalization , Pandemics , Pneumonia, Viral , Diagnosis , Retrospective StudiesABSTRACT
Objective To explore molecular mechanisms of dexmedetomidine(Dex)function as protec-tive effect on acute lung injury(ALI). Methods Hemorrhagic shock ALI rat model was constructed and divided into 6 groups at random:C group,control group;A group,ALI;G,ALI rat groups were treated with 5μg/kg;I1 group,ALI+PDTC(200 mg/kg);I2 group,ALI+STA-21(100 mg/kg). After 12 h modeled,all rats were anes-thetized and killed,and BALF was collected and the concentrations of TNF-α,IL-6,IL-1β,IL-4 were deter-mined using ELISA. HE staining to observe the morphology of lung tissue. Expression of NF-κB,STAT-3,p-STAT-3 proteins were determined using immunohistochemistry or Western blot. Results In A group,the structure of lung tissue were strongly destroyed ,a large number of neutrophils invasion and red blood cells leakage was found;lung tissue structure in G,I1 and I2 groups were gross integrity. The concentrations of TNF-α,IL-6,IL-1β,IL-4 in A group were significantly higher than C group(P<0.05). The concentrations of TNF-α,IL-6,IL-1β,IL-4 in G group,I1 group and I2 group was significantly lower than A group(P<0.05). The levels of NF-κB and p-STAT-3 protein in A group were obvious enhanced(P < 0.05)compared with G group,I1 group,I2 group,respectively. Conclusion Dex inhibited inflammatory to exert lung protective effect via inhibiting NF-κB,STAT-3 activation-induced by ALI.
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Objective To explore molecular mechanisms of dexmedetomidine(Dex)function as protec-tive effect on acute lung injury(ALI). Methods Hemorrhagic shock ALI rat model was constructed and divided into 6 groups at random:C group,control group;A group,ALI;G,ALI rat groups were treated with 5μg/kg;I1 group,ALI+PDTC(200 mg/kg);I2 group,ALI+STA-21(100 mg/kg). After 12 h modeled,all rats were anes-thetized and killed,and BALF was collected and the concentrations of TNF-α,IL-6,IL-1β,IL-4 were deter-mined using ELISA. HE staining to observe the morphology of lung tissue. Expression of NF-κB,STAT-3,p-STAT-3 proteins were determined using immunohistochemistry or Western blot. Results In A group,the structure of lung tissue were strongly destroyed ,a large number of neutrophils invasion and red blood cells leakage was found;lung tissue structure in G,I1 and I2 groups were gross integrity. The concentrations of TNF-α,IL-6,IL-1β,IL-4 in A group were significantly higher than C group(P<0.05). The concentrations of TNF-α,IL-6,IL-1β,IL-4 in G group,I1 group and I2 group was significantly lower than A group(P<0.05). The levels of NF-κB and p-STAT-3 protein in A group were obvious enhanced(P < 0.05)compared with G group,I1 group,I2 group,respectively. Conclusion Dex inhibited inflammatory to exert lung protective effect via inhibiting NF-κB,STAT-3 activation-induced by ALI.
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L-tryptophan, one of the aromatic amino acids, is widely used in the fields of medicine, food and feed additives. The phosphoenolpyruvate-carbohydrate phosphotransferase system (PTS) plays an important role in glucose transport and phosphorylation in Escherichia coli. PTS-mediated regulation dominates the carbohydrates' uptake and metabolism in E. coli. We constructed L-tryptophan-producing bacteria containing two typical PTS mutations (ptsHIcrrglf⁻ glk⁺ and ptsG⁻) by Red homologous recombination system, and studied in 50 L jar fermenter using fed-batch fermentation. Both PTS system mutants had a great impact on the biomass (increasing 47.0% and 17.6%, respectively), L-tryptophan production (increasing 25.9% and 9.4%, respectively), glucose conversion rate (increasing 26.5% and 17.4%, respectively) and byproduct acetic acid generation (slightly increased and decreased，respectively).
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Objective To evaluate the effect of dexmedetomidine on the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) during focal cerebral ischemia-reperfusion (I/R) in rats.Methods Thirty pathogen-free male Sprague-Dawley rats,aged 12-16 months,weighing 300-360 g,were randomly divided into 3 groups (n =10 each) using a random number table:sham operation group (group Sham),focal cerebral I/R group (group I/R),and dexmedetomidine group (group D).Focal cerebral I/R was induced by occlusion of the right middle cerebral artery.In group D,dexmedetomidine was given as a loading dose of 1 μg/kg (over 10 min) starting from 1 h of ischemia,followed by an infusion of 0.05 μg · kg-1 · h-1 until 2 h of reperfusion.Neurological deficit was assessed and scored at 24 h of reperfusion,and then the rats were sacrificed.Brains were removed for determination of cerebral infarct size and expression of iNOS and COX-2 in the hippocampus (by Western blot).The percentage of cerebral infarct size was calculated.Results Compared with group Sham,the neurological deficit score,percentage of head swing to the left,percentage of cerebral infarct size,and expression of iNOS and COX-2 in the hippocampus were significantly increased in I/R and D groups (P＜0.05).Compared with group I/R,the neurological deficit score,percentage of head swing to the left,percentage of cerebral infarct size,and expression of iNOS and COX-2 in the hippocampus were significantly decreased in group D (P＜0.05).Conclusion The mechanism by which dexmedetomidine mitigates focal cerebral I/R injury may be related to inhibition of iNOS and COX-2 expression in rats.
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promoted by high glucose, which can enlarge the biological effect of PAF.
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Objective To study the effect of high glucose and lysophosphatidylcholine (LPC) on the role of fibronectin and plate-let activating factor (PAF) via the interaction of endothelial cells and mesangial cells. Methods The model of intercellular interaction be-tween endothelial cells and mesangial cells was established and divided into 4 groups: control, mannitol, high glucose and LPC, and BN52021 group. The level of fibronectin and PAF were determined by enzyme linked immunosorbent assay in the culture media. Results The level of fibronectin and PAF of high glucose and LPC group were higher than those of control group in co-culture and monolayer cell cul-ture (P<0.05). Intervened by high glucose and LPC, the level of fibronectin and PAF of co-culture were higher than those from monolay-er cell culture (P<0.05). The level of fibrocentin in the BN52021 group was lower than that of high glucose and LPC (P<0.05). Con-clusions Exposed to high glucose and LPC, endothelial cells and mesangial cells can interact with each other to produce more fibrocentin and PAF. The increase of fibronectin is partly concerned with PAF.
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Objective To study the effect of astragalus membranaceus on the interaction of mesangial cells and endothelial cells in the media of high glucose and lysophosphatidylcholine. Methods The model of intercellular interaction between endothelial cells and mesangial cells of diabetic nephropathy was established and divided into 4 groups:control, mannitol, high glucose and lysophosphatidylcholine, intervented with astragalus membranaceus. Endothelial cells and mesangial cells were co-cultured in DMEN with or without astragalus membranaceus in high glucose and lysophosphatidylcholine media up to 24 hours. The level of collagen Ⅳ and fibrocentin were determined by enzyme linked immunosorbent assay in the culture media. Results The level of collagen Ⅳ and fibrocentin of high glucose and lysophosphatidylcholine group was markedly higher than that of control group in co-culture and the ordinary monolayer cell culture (P